豚鼠腹腔巨噬细胞的分离培养及免疫抗结核功能检测

巨噬细胞（macrophages，Mφ）在机体正常发育、体内平衡、组织修复和对病原体的免疫反应中起着重要作用。为了研究豚鼠（Cavia porcellus）Mφ在结核免疫中的作用及机制，本研究通过淀粉肉汤溶液预刺激的方法分离培养了豚鼠腹腔Mφ，并进行了细胞形态学观察、细胞活力、吞噬功能、非特异性酯酶染色和细胞表面特异性抗原表达检测等一系列鉴定。在此基础上，利用结核分枝杆菌（Mycobacteriumtuberculosis，MTB）卡介苗（BacillusCalmette—Gu6rin，BCG）感染分离培养的Mqo，并分析BCG对Mφ NO的产生和细胞凋亡的影响。结果表明，分离培养的腹腔Mφ具...

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Bibliographic Details
Published in农业生物技术学报 Vol. 25; no. 8; pp. 1307 - 1313
Main Author 李欧阳 洪丹萍 柯敏霞 麦丽丽 沈文强 何玉龙 刘立丽 吴月红
Format Journal Article
LanguageChinese
Published 2017
Subjects
NO
凋亡
卡介苗（BCG）
巨噬细胞（Mφ）
豚鼠
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Summary:巨噬细胞（macrophages，Mφ）在机体正常发育、体内平衡、组织修复和对病原体的免疫反应中起着重要作用。为了研究豚鼠（Cavia porcellus）Mφ在结核免疫中的作用及机制，本研究通过淀粉肉汤溶液预刺激的方法分离培养了豚鼠腹腔Mφ，并进行了细胞形态学观察、细胞活力、吞噬功能、非特异性酯酶染色和细胞表面特异性抗原表达检测等一系列鉴定。在此基础上，利用结核分枝杆菌（Mycobacteriumtuberculosis，MTB）卡介苗（BacillusCalmette—Gu6rin，BCG）感染分离培养的Mqo，并分析BCG对Mφ NO的产生和细胞凋亡的影响。结果表明，分离培养的腹腔Mφ具有Mφ所特有的形态特征、较强细胞活力和吞噬能力，并可检测到特异性的白细胞分化抗原14（cluster of differentiation14，CD14）、CD40和CD68等的表达。与未感染的对照组相比，BCG感染24h后Mφ产生的NO极显著增加（P〈0．01），细胞的凋亡率也极显著升高（P〈0．01）。表明淀粉肉汤溶液刺激的方法可成功获取豚鼠腹腔Mφ，同时证实BCG感染后能诱导其产生的NO增多，并且细胞凋亡率增加，提示其可能在抗结核免疫中起重要作用。研究结果为MTB和Mφ相互作用研究提供细胞模型和数据支撑。
Bibliography:11-3342/S
Guinea pig, Macrophage （Mφ）, Bacillus Calmette-Guerin （BCG）, NO, Apoptosis
Macrophages are key cells associated with innate immunity, pathogen containment and modulation of the immune response and also the key regulators of tissue repair, regeneration and fibrosis. Commonly used model systems for studying macrophage biology have centered on macrophage-like leukemic cell lines, primary macrophages derived from model organisms and primary macrophages differentiated from blood monocytes. Although these cells have provided important insights into macrophage-associated biology, there are issues that need consideration. The primary drawback of understanding macrophage biology is that reliable and scalable macrophage models for cellular and genetic studies is scarce, limiting their utility in genetic studies. Based on these, in order to elucidate the role and mechanism of guinea pig macrophages in tuberculosis immunization, in the present study, guinea pig peritoneal macrophages were isolated and characteriz
ISSN:1674-7968